Fertility and Sterility - Articles in Press

Effect of 5-aza-2′-deoxycytidine on methylation of the putative imprinted control region of H19 during the in vitro development of vitrified bovine two-cell embryos - Corrected Proof

2012-05-23

Objective: To determine the effect of vitrification and 5-aza-2′-deoxycytidine (5-aza-dC) on the methylation levels of the putative imprinted control region (ICR) of H19 and H19 expression in bovine two-cell embryos and their derived blastocysts.Design: Experimental animal study.Setting: Academic institution.Animal(s): Abattoir-derived bovine ovaries.Intervention(s): Vitrified two-cell embryos were cultured in vitro to blastocysts with 0.01 μM 5-aza-dC (5-aza-dC group) or without 5-aza-dC (vitrification group). Fresh embryos and their derived blastocysts were used as control.Main Outcome Measure(s): Putative ICR methylation of H19 was measured by bisulfate mutagenesis and sequencing, blastocyst development rate; total cell number were determined; and H19 expression was measured by real-time reverse transcriptase-polymerase chain reaction (PCR).Result(s): Vitrification significantly increased putative ICR methylation of H19 in two-cell embryos and their derived blastocysts; 5-aza-dC significantly reduced putative ICR methylation of H19 in vitrified two-cell embryos and their derived blastocysts. The H19 expression level was significantly higher in blastocysts from the 5-aza-dC group than the vitrification group. The blastocyst development rate and total cell number in the 5-aza-dC and vitrification groups were similar.Conclusion(s): Putative ICR methylation levels of H19 significantly increased in vitrified two-cell embryos and their derived blastocysts; 5-aza-dC significantly reduced putative ICR methylation of H19 and increased H19 expression in blastocysts derived from vitrified two-cell embryos.

Thyroid peroxidase antibody in women with unexplained recurrent miscarriage: prevalence, prognostic value, and response to empirical thyroxine therapy - Corrected Proof

2012-05-23

Objective: To determine the prevalence, prognostic value, and response to thyroxine therapy of thyroid peroxidase antibody (TPOAb) in women with unexplained recurrent miscarriage (RM).Design: Observational, cohort study. The index cases included women with unexplained RM who tested positive for thyroid peroxidase antibody, and control cases included women with unexplained RM who tested negative for the antibodies; a second age-matched control group included women with RM who had a known cause for the repeated pregnancy loss.Setting: Tertiary referral center for RM.Patient(s): A total of 496 women with unexplained RM and 220 women with known diagnoses of RM who had a TPOAb test.Intervention(s): Thyroxine replacement (50 μg daily during pregnancy) was begun in some patients who tested positive for thyroid peroxidase antibody, irrespective of TSH level.Main Outcome Measure(s): Miscarriage and live birth rates of a subsequent pregnancy.Result(s): A total of 496 women with unexplained RM who had a TPOAb test were included in the study. Of these, 10.7% of subjects tested positive for TPOAb. The prevalence of TPOAb in control subjects who had a known cause for RM was 11.8%. The live birth rate of the first pregnancies after referral was 64%, 53%, and 58% in TPOAb-negative, TPOAb-positive with thyroxine treatment, and TPOAb-positive without treatment subjects; there was no significant difference in the outcome between any two or three groups, or between those who tested positive or negative for TPOAb. Among women who tested positive for TPOAb, there was no difference in the antibody titer between women with unexplained RM and those with a known cause for the pregnancy loss. Women who tested positive for TPOAb were significantly more likely to have TSH levels above the normal range (≥4.2 mIU/L).Conclusion(s): The prevalence of TPOAb-positive results in women with unexplained RM is not higher than in the general population, TPOAb-positive status does not have a prognostic value regarding the outcome of a subsequent pregnancy, and empirical thyroxine therapy in those who tested positive did not seem to improve outcome.

Paternal and maternal carriage of the annexin A5 M2 haplotype are equal risk factors for recurrent pregnancy loss: a pilot study - Corrected Proof

2012-05-23

Objective: To study the possible contribution of paternal, in addition to maternal, carriage of M2/ANXA5 as a risk factor for recurrent pregnancy loss (RPL).Design: Case–control study.Setting: Academic research center.Patient(s): Couples presenting themselves to the Fertility Center, Ludwig-Maximilians-University Munich with two or more consecutive, unexplained miscarriages were selected for this study. Fertile female controls were from the same center and also from the resource of the Institute of Human Genetics, Westfalian Wilhelms-University Muenster. Population controls were drafted from the PopGen biobank, University Clinic Schleswig-Holstein Kiel.Intervention(s): None.Main Outcome Measure(s): Incidence of M2 carriage was estimated in patient and control groups, odds ratios were calculated, and RPL risk was evaluated.Result(s): In comparison with female fertile controls, the risk for repeated abortion in the RPL group, associated with M2 carriage, was between 1.7 and 3.8, and it was 2.3 compared with population controls. Because of the equal genetic incidence of M2, with an allelic frequency of 0.167 in the female and male partner RPL subgroups, the haplotype confers approximately the same relative risk to carriers of both sexes.Conclusion(s): Paternal M2 carriage seems to confer an equal risk for recurrent miscarriages as M2 carriage in RPL mothers. This finding points to a role of ANXA5 and the M2 haplotype in the fetus and/or the extraembryonic membranes for pregnancy pathology. Prognostic RPL algorithms might be improved by testing the male partner for M2 carriage, and this may guide adequate therapeutic decisions.

Human sperm head vacuoles are physiological structures formed during the sperm development and maturation process - Corrected Proof

Atsushi Tanaka, Motoi Nagayoshi, Izumi Tanaka, Hiroshi Kusunoki — 2012-05-23

Objective: To clarify whether human sperm vacuoles affected intracytoplasmic sperm injection (ICSI) success rates.Design: Retrospective study.Setting: A private infertility clinic.Patient(s): Spermatozoa and spermatids were obtained from 11 normozoospermic, 10 oligozoospermic or asthenozoospermic, 4 obstructive azoospermic, and 3 nonobstructive azoospermic men.Intervention(s): Differential interference contrast observation and intracytoplasmic injection of morphologically selected sperm.Main Outcome Measure(s): Incidence, size, and position of vacuoles of sperm cells were recorded. Ability of fertilization and blastocyst development were compared between cells with and without vacuoles.Result(s): More than 97.4% of ejaculated, 87.5% of epididymal, 87.5% of testicular spermatozoa, and more than 90.0% of Sc-Sd2 spermatids had vacuoles of various sizes. The incidence of vacuoles on ejaculated cells was significantly higher than that on the other types of cells, but there was no difference between sperm from normozoospermic men and those from the other donors. Removal of plasma membrane and/or acrosome did not affect the incidence of vacuoles. Although more than 60% of spermatozoa had small vacuoles in the acrosomal regions, 52.6% of Sb1-2 spermatids had large vacuoles. After injection of a motile spermatozoon with large and small vacuoles, 60.9% and 85.7% of metaphase II oocytes could be normally fertilized, respectively, and almost half of the zygotes developed to the blastocyst stage. When using sperm without vacuoles, the fertilization rate was 80.0%, but only 25% of them developed to the blastocyst stage.Conclusion(s): Human sperm head vacuoles did not affect ICSI outcomes.

Tetraspanin-interacting protein IGSF8 is dispensable for mouse fertility - Corrected Proof

Naokazu Inoue, Takao Nishikawa, Masahito Ikawa, Masaru Okabe — 2012-05-21

Objective: To determine the physiological role of IGSF8 for fertility.Design: Experimental prospective study.Setting: Academic basic research laboratory.Animal(s): C57BL/6J and hybrid B6D2F1 mice, as well as Cd9 and Igsf8 knockout mice (C57BL/6J and 129/SvJ mix background), were used for this study.Intervention(s): None.Main Outcome Measure(s): In vitro and in vivo fertility tests of Igsf8 knockout mice.Result(s): Tetraspanin family member CD9 plays an important role in sperm-egg fusion. Recently, some researchers have reported that CD9 tightly associates with the immunoglobulin superfamily member IGSF8 on the egg surface and that IGSF8 is undetectable on the surface of Cd9-deficient eggs. This led us to hypothesize that IGSF8 participates in sperm-egg fusion together with CD9. To examine the physiological role of IGSF8 in vivo, we generated Igsf8-deficient mice by homologous recombination and examined the fertility of the females.Conclusion(s): The Igsf8-deficient female mice showed no fertilization defect in vitro or in vivo. We observed that Igsf8-deficient eggs retained the normal level and localization of CD9, resulting in normal microvilli formation, which indicates that IGSF8 is dispensable in fertility.

Maternal characteristics and twin gestation outcomes over 10 years: impact of conception methods - Corrected Proof

2012-05-18

Objective: To compare maternal characteristics and obstetric outcomes of spontaneously conceived (SC) and after–fertility treatment (FT) twins.Design: Retrospective study.Setting: Single tertiary center (university hospital).Patient(s): All twin pregnancies (n = 1,239) delivered ≥24 weeks of gestation and classified by the mode of conception.Intervention(s): None.Main Outcome Measure(s): Maternal age, prematurity, preterm rupture of membranes (PROM), delivery mode, cervical insufficiency, preeclampsia/HELLP (hemolysis, elevated liver enzymes, low platelet count) syndrome, gestational diabetes, and postpartum hemorrhage.Result(s): Of 1,239 mothers evaluated, 34.4% received FT. We observed a continuous 3.2-year mean increase in maternal age over time in the FT population. The FT group had statistically significantly higher rates of women ≥35 years of age versus the SC group (37.6% vs. 22.9%). Mean maternal age (32.5 vs. 30.1 years) and the rates of cesarean delivery (72.3% vs. 63.9%), dichorionic twin prematurity (67.7% vs. 59.6%), postpartum hemorrhage (9.9% vs. 6%), PROM (2.3% vs. 0.6%), and cervical insufficiency (17.6% vs. 10%) were statistically significantly higher in the FT group. Preeclampsia/HELLP syndrome and gestational diabetes showed no significantly significant difference.Conclusion(s): Mothers of twins after FT were statistically significantly older and had higher rates of prematurity, cesarean delivery, and obstetric morbidity.

Metformin: direct inhibition of rat ovarian theca-interstitial cell proliferation - Corrected Proof

2012-05-18

Objective: To determine whether metformin has direct effects on ovarian theca-interstitial (T-I) cell proliferation through activation of adenosine monophosphate–activated protein kinase (AMPK).Design: In vitro experimental study.Setting: Academic medical center laboratory.Animal(s): Immature Sprague-Dawley female rats.Intervention(s): Ovarian T-I cells were isolated, purified, and cultured in the absence (control) or presence of insulin (1 μg/mL) with or without metformin or other activators/inhibitors of AMPK (AICAR, compound C).Main Outcome Measure(s): Proliferation assessed by determination of expression levels of proteins involved in cell cycle progression, cyclin D3, and cyclin-dependent kinase 4 (CDK4) with Western blot analysis, and determination of DNA synthesis with bromodeoxyuridine (BrdU) incorporation assay; activation of AMPK, Erk1/2, and S6K1 determined by Western blot analysis with the use of antibodies specific for the phosphorylated (activated) forms.Result(s): Metformin inhibited insulin-induced ovarian T-I cell proliferation and the up-regulation of the cell cycle regulatory proteins, cyclin D3 and CDK4. Metformin independently activated AMPK in a dose-dependent manner. Treatment with metformin inhibited insulin-induced activation of Erk1/2 and S6K1. This effect was reversed with the addition of compound C, a known AMPK inhibitor.Conclusion(s): Metformin directly inhibits proliferation of ovarian T-I cells via an AMPK-dependent mechanism. These findings further validate the potential benefits of metformin in the treatment of conditions associated with hyperinsulinemia and excessive growth of ovarian T-I cells (such as polycystic ovary syndrome).

Inhibitory effects of methotrexate on spontaneous motility and Cajal-like type of tubal interstitial cells in rabbit oviduct - Corrected Proof

Xiao-Jun Yang, Wei Wei, Jing Zhao, Fei-Yun Zheng — 2012-05-18

Objective: To study the adverse biomechanical effects of methotrexate (MTX) on spontaneous tubal motility and on a widely distributed Cajal-like type of tubal interstitial cells (t-ICC) in rabbits. In our previous study, MTX was confirmed to cause acute endosalpingitis, and ultrastructural and steroid receptor damage in rat's endosalpinx in a dose-dependent manner.Design: Differences in spontaneous tubal contractions and cellular distribution of t-ICC in isthmus were evaluated in response to MTX.Setting: Medical school research laboratory.Animal(s): Twenty nonpregnant female New Zealand albino rabbits in estrus stage were divided equally into four groups.Intervention(s): Rabbits received IM MTX (1, 5, 10 mg/kg body weight) and controls received physiological saline.Main Outcome Measure(s): On day 7, in vitro motility studies measuring spontaneous tubal contractions were performed, and cellular distribution of t-ICC was determined by immunohistochemistry.Result(s): Methotrexate produced a concentration-dependent inhibition of spontaneous isthmus contractions (frequency in 5, 10 mg/kg groups, and amplitude in 1, 5, 10 mg/kg MTX groups). It decreased significantly compared with the control group. Meanwhile, MTX at 5, 10 mg/kg decreased the population of c-kit immunoreactive t-ICC significantly.Conclusion(s): The decreased t-ICC may contribute to the diminished tubal smooth muscle contractility caused by MTX as observed. Tubal interstitial cells might be new potential targets for a variety of dysfunctional tubal motility diseases.

Is the zona pellucida thickness of human embryos influenced by women's age and hormonal levels? - Corrected Proof

2012-05-18

Objective: To evaluate whether zona pellucida thickness (ZPT) of human embryos is correlated with maternal age, patient's hormonal status, embryo quality, and IVF outcomes.Design: Prospective study.Setting: University-affiliated IVF clinic.Patient(s): Couples undergoing IVF-ET cycles.Intervention(s): Zona measurements, clinical data collection.Main Outcome Measure(s): Correlation between the ZPT and maternal age, basal FSH and E2 levels, stimulation protocols, cause of infertility, embryo quality, and implantation/pregnancy rates.Result(s): The measurements of ZPT were collected from 5,184 day 3 human embryos originated from 744 IVF patients. The overall mean ZPT was 16.18 ± 2.00 μm. No significant correlation was observed between the ZPT and the patient's age, E2 values on the day of hCG administration, basal concentration of serum FSH, stimulation protocol, infertility diagnosis, and implantation/pregnancy rates. The ZPT was strongly influenced only by the embryo quality: Embryos with good morphology exhibited considerably thinner ZP compared with those of less favorable morphology (mean 15.87 ± 2.48 μm vs. 16.36 ± 2.57 μm, respectively). The ZPT had no significant impact on the implantation and pregnancy rates.Conclusion(s): The thickness of the human ZP of day 3 embryos is not influenced by women's age and hormonal levels. The strong correlation between ZPT and embryo quality suggests that thickness of ZP depends on inherent embryo properties. The overall ZPT is not a good predictive indicator for IVF clinical outcomes.

Expression of phosphoinositide-specific phospholipase C enzymes in normal endometrium and in endometriosis - Corrected Proof

2012-05-18

Objective: To delineate the panel of expression of phosphoinositide-specific phospholipase C (PI-PLC) signaling enzymes in normal endometrium and in endometriosis.Design: Clinical/experimental study.Setting: University.Patient(s): Healthy donor woman and endometriosis-affected woman.Intervention(s): Normal endometrium and endometriosis surgical biopsies were analyzed using gene expression analyses methodology (reverse transcriptase-polymerase chain reaction [PCR], bioanalyses).Main Outcome Measure(s): Gene expression (messenger RNA concentration) measures of 12 PI-PLC enzymes: PI-PLC β1, PI-PLC β2, PI-PLC β3, PI-PLC β4, PI-PLC γ1, PI-PLC γ2, PI-PLC δ1, PI-PLC δ3, PI-PLC δ4, PI-PLC ε, PI-PLC η1, and PI-PLC η2.Result(s): PI-PLC β1, PI-PLC β3, PI-PLC δ1, and PI-PLC δ3 enzymes were detected, although differently expressed in normal and endometriosis tissues.Conclusion(s): The involvement of PI-PLC enzymes in inflammation and the consistency of susceptible endometriosis loci with PI-PLC genes mapping corroborate the hypothesis that PI signaling might be involved in the pathogenesis of endometriosis.

Spermatogonial stem cell preservation in boys with Klinefelter syndrome: to bank or not to bank, that's the question - Corrected Proof

2012-05-18

Although early development of testis appears normal in boys with Klinefelter syndrome (KS), spermatogonial stem cell (SSC) depletion occurs in midpuberty, leading to infertility. Therefore, freezing of semen samples or testicular tissue sampling could be offered to boys with KS at onset of puberty. However, only in about half of patients with KS, adult or prepubertal, spermatozoa or SSCs can be observed, and to date, no clinical parameters are available to detect patients who might benefit from these techniques. Furthermore, strategies for the further use of the cryopreserved material are still under investigation. Retrieval of spermatogonial cells in prepubertal boys with KS should therefore still be viewed as experimental and patients and their parents must be counseled accordingly.

Oocyte vitrification does not increase the risk of embryonic aneuploidy or diminish the implantation potential of blastocysts created after intracytoplasmic sperm injection: a novel, paired randomized controlled trial using DNA fingerprinting - Corrected Proof

2012-05-18

Objective: To assess the impact of oocyte vitrification on aneuploidy and reproductive potential by comparing vitrified and control oocytes from a single patient within a single cycle and a single fresh transfer.Design: Paired randomized controlled trial in which each patient's cohort of mature oocytes was divided into two even groups with half undergoing Cryotop vitrification and rapid warming and half serving as controls.Setting: Academic center for reproductive medicine.Patient(s): Forty-four patients with a mean age of 29.9 ± 2.3 years and normal ovarian reserve.Intervention(s): Cryotop vitrification of half of mature oocytes. Trophectoderm biopsy with single nucleotide polymorphism microarray analysis for ploidy and DNA fingerprinting.Main Outcome Measure(s): Rate of aneuploidy (primary outcome), fertilization, cleavage, blastulation, and implantation in embryos derived from vitrified and control oocytes.Result(s): A total of 588 mature oocytes were randomized, with 240/294 (81.6%) surviving vitrification. Among surviving vitrified oocytes, there was a lower fertilization rate with intracytoplasmic sperm injection (77.9% vs. 90.5%; relative risk [RR], 0.86; 95% confidence interval [CI], 0.80–0.93), a lower cleavage rate (90.9% vs. 99.2%; RR, 0.92; 95% CI, 0.87–0.96), and a lower usable blastocyst formation rate per two pronuclei (34.8% vs. 50.8%; RR, 0.68; 95% CI, 0.54–0.86). There was no difference in the rate of embryonic aneuploidy (vitrified, 29.1% vs. control, 26.4%). In paired blastocyst transfers, the ongoing pregnancy rate per embryo transferred was similar (vitrified, 53.9% vs. control, 57.7%).Conclusion(s): Although the IVF process is less efficient after oocyte vitrification, implantation rates are equivalent and there is no increased risk of aneuploidy. Given the lack of other viable options, this study provides great reassurance to patients and clinicians applying oocyte vitrification for fertility preservation.

Frozen-thawed spermatozoa from oligozoospermic ejaculates are susceptible to in situ DNA fragmentation in polyvinylpyrrolidone-based sperm-immobilization medium - Corrected Proof

2012-05-18

Objective: To elucidate the effect of sperm immobilization media that are and are not based on polyvinylpyrrolidone (PVP) on the DNA integrity of fresh and frozen-thawed spermatozoa during standard intracytoplasmic sperm injection (ICSI) conditions.Design: Experimental prospective study.Setting: Embryology research laboratory.Patient(s): Forty-six ejaculates from normozoospermic and oligozoospermic men.Intervention(s): Assessment of sperm DNA fragmentation by single-cell gel electrophoresis assay.Main Outcome Measure(s): DNA integrity of fresh and frozen-thawed spermatozoa from normozoospermic and oligozoospermic ejaculates exposed to PVP-based and non-PVP-based media.Result(s): Exposure of fresh and frozen thawed spermatozoa from normozoospermic and oligozoospermic ejaculates to PVP-based medium in an ICSI dish for 30 minutes statistically significantly increased the DNA fragmentation. In contrast, the extent of DNA fragmentation in non-PVP-based medium did not statistically significantly differ from control.Conclusion(s): A PVP-based medium can induce a statistically significant amount of sperm DNA fragmentation in an ICSI dish, and frozen-thawed sperm from oligozoospermic ejaculates are more susceptible to in situ DNA fragmentation.

Rescue human chorionic gonadotropin for false empty follicle syndrome: optimism for successful pregnancy outcome - Corrected Proof

2012-05-18

Objective: To describe two cases of successful pregnancy after a rescue course of hCG in the setting of false empty follicle syndrome.Design: Case report.Setting: Academic medical center.Patient(s): Two patients undergoing ultrasound-guided oocyte retrieval with failure to obtain oocytes during oocyte retrieval.Intervention(s): Rescue course of hCG with second oocyte retrieval 35 hours later.Main Outcome Measure(s): Live birth.Result(s): Two live-birth pregnancies.Conclusion(s): Live-birth pregnancies are a realistic possibility after administration of a rescue course of hCG and repeat oocyte retrieval in the setting of false empty follicle syndrome.

Adipose expression of adipocytokines in women with polycystic ovary syndrome - Corrected Proof

2012-05-17

Objective: To investigate the role of adipocytokines in the pathophysiology of polycystic ovary syndrome (PCOS) by analyzing the messenger RNA (mRNA) expression and plasma levels of adipocytokines.Design: Cross sectional study.Setting: Hospital.Patient(s): Thirty-six women with PCOS, 17 lean (LP) and 19 obese (OP), and 24 age- and weight-matched controls, 8 lean (LC) and 16 obese (OC).Intervention(s): Subcutaneous adipose tissue and fasting plasma samples collected from 60 women, and insulin sensitivity evaluated by euglycemic hyperinsulinemic clamp and homeostatic model assessment insulin resistance index (HOMA-IR).Main Outcome Measure(s): mRNA expression of adiponectin, leptin, and interleukin-6 (IL-6) in adipose tissue, and plasma levels of leptin, adiponectin, resistin, visfatin, and tumor necrosis factor α (TNF-α).Result(s): The baseline data on body mass index (BMI), age, androgen levels, and insulin sensitivity was published previously. We found no independent effect of PCOS on the adipose expression of leptin, adiponectin, or IL-6 or on the plasma levels of adiponectin, leptin, resistin, visfatin, and TNF-α. Obesity was associated with increased mRNA expression of leptin, lower expression of adiponectin, and increased plasma levels of leptin.Conclusion(s): Obesity is per se associated with increased adipose expression and plasma levels of leptin, lower expression of adiponectin, and marginally elevated expression of IL-6, but PCOS does not appear to have an independent effect on the adipose expression of leptin, adiponectin, and IL-6 or the circulating adipocytokines.Clinical Trial Registration Number: NCT00975832.

Animal models of polycystic ovary syndrome: a focused review of rodent models in relationship to clinical phenotypes and cardiometabolic risk - Corrected Proof

Danni Shi, Donna F. Vine — 2012-05-17

Objective: To review rodent animal models of polycystic ovary syndrome (PCOS), with a focus on those associated with the metabolic syndrome and cardiovascular disease risk factors.Design: Review.Animal(s): Rodent models of PCOS.Intervention(s): Description and comparison of animal models.Main Outcome Measure(s): Comparison of animal models to clinical phenotypes of PCOS.Result(s): Animals used to study PCOS include rodents, mice, rhesus monkeys, and ewes. Major methods to induce PCOS in these models include subcutaneous injection or implantation of androgens, estrogens, antiprogesterone, letrozole, prenatal exposure to excess androgens, and exposure to constant light. In addition, transgenic mice models and spontaneous PCOS-like rodent models have also been developed.Conclusion(s): Rodents are the most economical and widely used animals to study PCOS and ovarian dysfunction. The model chosen to study the development of PCOS and other metabolic parameters remains dependent on the specific etiologic hypotheses being investigated. Rodent models have been shown to demonstrate changes in insulin metabolism, with or without induction of hyperandrogenemia, and limited studies have investigated cardiometabolic risk factors for type 2 diabetes and cardiovascular disease. Given the clinical heterogeneity of PCOS, the utilization of different animal models may be the best approach to further our understanding of the pathophysiologic mechanisms associated with the early etiology of PCOS and cardiometabolic risk.

Body mass index and short-term weight change in relation to treatment outcomes in women undergoing assisted reproduction - Corrected Proof

2012-05-17

Objective: To assess the relation between body mass index (BMI) and short-term weight change with assisted reproductive technology (ART) outcomes.Design: Prospective cohort study.Setting: Fertility center.Patient(s): A total of 170 women undergoing 233 ART cycles.Intervention(s): Baseline BMI and short-term weight change were related to ART outcomes. Regression models accounting for repeated observations were used to adjust data for potential confounders.Main Outcome Measure(s): Peak E2 levels, oocyte yield, MII yield, fertilization rate, embryo quality, postive [beta]-hCH, clinical pregnancy and live birth rates.Result(s): Overweight and obesity were associated with lower live birth rates. The adjusted live birth rate (95% confidence interval) was 42% (28%–58%) among women with a BMI between 20 and 22.4 kg/m2 and 23% (14%–36%) among overweight or obese women. Short-term weight loss was associated with a higher proportion of metaphase II (MII) oocytes retrieved. The adjusted proportion of MII eggs was 91% (87%–94%) for women who lost 3 kg or more and 86% (81%–89%) for women whose weight remained stable. This association was stronger among women who were overweight or obese at baseline. Short-term weight loss was unrelated to positive β-hCG, clinical pregnancy, or live birth rates.Conclusion(s): Overweight and obesity were related to lower live birth rates in women undergoing ART. Short-term weight loss was related to higher MII yield, particularly among overweight and obese women, but unrelated to clinical outcomes.

Reduced pretreatment ovarian reserve in premenopausal female patients with Hodgkin lymphoma or non-Hodgkin-lymphoma—evaluation by using antimüllerian hormone and retrieved oocytes - Corrected Proof

2012-05-17

Objective: To study whether the ovarian reserve in female lymphoma patients is already reduced before the start of chemotherapy.Design: Age-matched control study.Setting: Women's university hospital.Patient(s): Female patients aged <40 years with the initial diagnosis of lymphoma (study group) were compared with age-matched healthy volunteers (control group). Eighty-four female patients with breast cancer and 64 patients with lymphoma who underwent ovarian hormonal stimulation as a fertility-preserving method before the start of chemotherapy.Intervention(s): Measurement of antimüllerian-hormone (AMH) levels. Ovarian hormonal stimulation to retrieve oocytes.Main Outcome Measure(s): AMH levels of the lymphoma patients and the healthy volunteers were compared. Numbers of retrieved oocytes after hormonal stimulation in patients with breast cancer and those with lymphoma were compared.Result(s): Female lymphoma patients have significantly lower AMH levels than healthy age-matched controls: mean value of AMH was 2.06 ng/mL in the study group versus 3.20 ng/mL in the control group. Analysis of the stimulation results showed that in significantly younger patients with lymphoma, significantly fewer oocytes could be retrieved in comparison to those with breast cancer.Conclusion(s): Ovarian reserve is reduced in female patients affected by lymphoma even before the start of chemotherapy. Proper counseling and implementation of fertility-preserving methods is highly recommended.

Integration of immunodeficiency virus in oocytes via intracytoplasmic injection: possible but extremely unlikely - Corrected Proof

2012-05-14

Objective: To determine if human oocytes can be infected with HIV-1 via intracytoplasmic injection and to determine the infection threshold.Design: Twenty-eight donated immature and unfertilized human oocytes from HIV-negative women were injected with 4 × 104 HIV-1 virions and 13 oocytes were used as uninjected controls. To determine the infection threshold, 543 cat oocytes were injected with 4 × 104, 4 × 102, or 40 copies of feline immunodeficiency virus (FIV) and 376 oocytes were used as controls.Setting: Academic hospital.Patient(s)/Animal(s): Donated immature human oocytes and mature cat oocytes.Intervention(s): Injection with HIV-1 or FIV.Main Outcome Measure(s): Viral integration as measured by fluorescent in situ hybridization with HIV-1-specific probes or by nested FIV polymerase chain reaction.Result(s): We detected viral integration in three of 28 (11%) human oocytes injected with 4 × 104 copies of HIV-1. When injected with high dose FIV (4 × 104 copies) 16%–49% of cat oocytes showed viral integration. This decreased to 2%–7% and 0.6%–1.8% when an intermediate (4 × 102 copies) or low (40 copies) dose was injected, respectively.Conclusion(s): Human and cat oocytes can be infected with HIV-1 and FIV respectively, when injected with high amounts of virus. The probability of viral integration is extremely low when small amounts of virus particles are injected. Taking into account the small volume injected during intracytoplasmic injection, the chances of viral integration are 0.00002%.

Pregnancies following ultrasound-guided drainage of tubo-ovarian abscess - Corrected Proof

2012-05-14

Objective: To study fertility among women treated by means of ultrasound-guided drainage and antibiotics for tubo-ovarian abscess (TOA).Design: Retrospective cohort study.Setting: A tertiary referral center.Patient(s): One hundred women of reproductive age treated for TOA between June 1986 and July 2003.Intervention(s): Transvaginal ultrasound-guided drainage of TOA was performed in all patients. The procedure was repeated if a substantial amount of pus was seen using ultrasonography 2–5 days after the initial aspiration, and repeated later if necessary.Main Outcome Measure(s): Frequency of naturally conceived pregnancies.Result(s): Twenty of 38 (52.6%; 95% CI 36.5–68.9%) women who intended to have a child achieved pregnancy naturally and became mothers. In addition, 7 (50%) of 14 women who were not on birth control on a regular basis became pregnant. No ectopic pregnancies were registered.Conclusion(s): Ultrasound-guided drainage of TOA in combination with antibiotics seems to preserve fertility in approximately half of the patients.

2-Methoxyestradiol causes functional repression of transforming growth factor β3 signaling by ameliorating Smad and non-Smad signaling pathways in immortalized uterine fibroid cells - Corrected Proof

Salama A. Salama, Concepcion R. Diaz-Arrastia, Gokhan S. Kilic, Marwa W. Kamel — 2012-05-14

Objective: To investigate the effects and the mechanism of action of 2-methoxyestradiol (2ME2) on transforming growth factor (TGF) β3–induced profibrotic response in immortalized human uterine fibroid smooth muscle (huLM) cells.Design: Laboratory study.Setting: University research laboratory.Patients(s): Not applicable.Interventions(s): Not applicable.Main Outcome Measure(s): huLM cells were treated with TGF-β3 (5 ηg/mL) in the presence or absence of specific Smad3 inhibitor SIS3 (1 μmol/L), inhibitor of the PI3K/Akt (LY294002, 10 μmol/L), or 2ME2 (0.5 μmol/L), and the expression of collagen (Col) type I(αI), Col III(αI), plasminogen activator inhibitor (PAI) 1, connective tissue growth factor (CTGF), and α-smooth muscle actin (α-SMA) were determined by real-time reverse-transcription polymerase chain reaction and immunoblotting. The effect of 2ME2 on Smad-microtubule binding was evaluated by coimmunoprecipitation.Result(s): Our data revealed that TGF-β3–induced fibrogenic response in huLM is mediated by both Smad-dependent and Smad-independent PI3K/Akt/mTOR signaling pathways. 2ME2 abrogates TGF-β3–induced expression of Col I(αI), Col III(αI), PAI-1, CTGF, and α-SMA. Molecularly, 2ME2 ameliorates TGF-β3–induced Smad2/3 phosphorylation and nuclear translocation. In addition, 2ME2 inhibits TGF-β3–induced activation of the PI3K/Akt/mTOR pathway.Conclusion(s): TGF-β3–induced profibrotic response in fibroid cells is mediated by Smad-dependent and Smad-independent PI3K/Akt/mTOR pathways. 2ME2 inhibits TGF-β3 profibrotic effects in huLM cells by ameliorating both Smad-dependent and Smad-independent signaling pathways.

Management of a cervical heterotopic pregnancy presenting with first-trimester bleeding: case report and review of the literature - Corrected Proof

Vasiliki A. Moragianni, Benjamin D. Hamar, Colin McArdle, David A. Ryley — 2012-05-14

Objective: To report a rare case of a cervical heterotopic pregnancy resulting from intrauterine insemination (IUI) that presented with first-trimester bleeding.Design: Case report and literature review.Setting: Large university-affiliated infertility practice.Patient(s): A 40-year-old gravida 2 para 1 Asian woman at 7-3/7 weeks gestational age following clomiphene citrate/IUI for the treatment of secondary infertility presented with heavy vaginal bleeding for several days.Intervention(s): Transvaginal ultrasound on admission revealed a single live intrauterine pregnancy and a cervical gestational sac containing a nonviable embryo. The patient continued to have vaginal bleeding and 2 days later underwent removal of the cervical ectopic pregnancy tissue with ring forceps, as well as an ultrasound-guided intracervical Foley balloon and cerclage placement. The bleeding subsided, and 48 hours later the Foley and cerclage were removed.Main Outcome Measure(s): Pregnancy outcome.Result(s): The remainder of the pregnancy was uncomplicated and the patient had a full-term cesarean delivery for footling breech of a healthy male infant.Conclusion(s): Cervical heterotopic pregnancy is a very rare event that almost universally results from infertility treatment. We present a case where we were able to remove the cervical ectopic and tamponade the bleeding, thus preserving the intrauterine pregnancy for this subfertile couple, and we review the existing literature.

The effect of body mass index on the outcomes of first assisted reproductive technology cycles - Corrected Proof

Vasiliki A. Moragianni, Stephanie-Marie L. Jones, David A. Ryley — 2012-05-14

Objective: To provide assisted reproductive technology (ART) outcome rates per body mass index (BMI) category after controlling for potential confounders.Design: Retrospective cohort study.Setting: Large university-affiliated infertility practice.Patient(s): Women undergoing ART.Intervention(s): None.Main Outcome Measure(s): The primary outcome was live birth. Analyses were stratified according to BMI category and adjusted for potential confounders, including maternal and paternal age, baseline serum FSH, duration of gonadotropin stimulation, mean daily gonadotropin dose, peak serum E2, number of oocytes retrieved, use of intracytoplasmic sperm injection, embryo quality and number, transfer day, and number of embryos transferred.Result(s): We analyzed the first autologous fresh IVF or IVF-ICSI cycle of 4,609 patients. There were no differences in the rates of cycle cancellation, spontaneous abortion, biochemical and ectopic pregnancies, or multiple births. After adjusting for potential confounders, patients with BMI ≥30.0 kg/m2 had significantly decreased odds of implantation, clinical pregnancy, and live birth. The adjusted odds ratio (95% confidence interval [CI]) of live birth were 0.63 (0.47–0.85) for BMI 30.00–34.99, 0.39 (0.25–0.61) for BMI 35.00–39.99, and 0.32 (0.16–0.64) for BMI ≥40.0 compared with normal-weight cohorts.Conclusion(s): Obesity has a significant negative effect on ART outcomes. Patients with BMI >30 kg/m2 have up to 68% lower odds of having a live birth following their first ART cycle compared with women with BMI <30.

Changes in hormonal profile and seminal parameters with use of aromatase inhibitors in management of infertile men with low testosterone to estradiol ratios - Corrected Proof

2012-05-14

Objective: To compare the effects of 2.5 mg letrozole with those of 1 mg anastrazole daily on the hormonal and semen profiles of a subset of infertile men with low T/E2 ratios.Design: Prospective, nonrandomized study.Setting: Reproductive medicine clinic.Patient(s): The study group consisted of 29 infertile men with a low serum T/E2 ratio (<10).Intervention(s): Patients were divided into two groups. Group A included 15 patients treated with 2.5 mg letrozole orally once daily for 6 months, and Group B consisted of 14 patients treated with 1 mg anastrazole orally every day for 6 months.Main Outcome Measure(s): Hormonal evaluation included measurement of serum FSH, LH, PRL, T, and E2. In all sperm analyses pretreatment and posttreatment total motile sperm counts (ejaculate volume × concentration × motile fraction) were evaluated.Result(s): The use of aromatase inhibitors (either letrozole or anastrazole) in cases of infertile men with low T/E2 ratios improved both hormonal and semen parameters.Conclusion(s): This study suggests that some men with severe oligospermia, low T levels, and normal gonadotropin concentration may have a treatable endocrinopathy.

Sirt1 exerts anti-inflammatory effects and promotes steroidogenesis in Leydig cells - Corrected Proof

2012-05-14

Objective: To investigate the anti-inflammatory effects of Sirt1 and its roles in steroidogenesis in Leydig cells.Design: In vitro cell culture study.Setting: Reproductive medical center.Animal(s): C57BL/6 male mice.Intervention(s): TM3 Leydig cells were treated with proinflammatory cytokines including tumor necrosis factor-α, interleukin-6, or interleukin-1β.Main Outcomes Measure(s): Sirt1 mRNA and protein levels were measured by real-time polymerase chain reaction and Western blotting in TM3 Leydig cells treated with proinflammatory cytokines. The cell viability was determined using MTT and BrdU incorporation assays. The effect of Sirt1 on the coactivation of steroidogenic factor 1 (SF-1) was characterized by coimmunoprecipitation and luciferase reporter assays.Result(s): Sirt1 mRNA and protein levels were significantly down-regulated by proinflammatory cytokines treatment in TM3 Leydig cells. Sirt1 agonist or overexpression could efficiently protect cytotoxicity induced by proinflammatory cytokines. Sirt1 promoted steroidogenesis through its coactivation of SF-1.Conclusion(s): Sirt1 plays protective roles and promotes steroidogenesis in Leydig cells through its anti-inflammatory actions and coactivation of SF-1.

Strain-specific spontaneous activation during mouse oocyte maturation - Corrected Proof

Yong Cheng, Zhisheng Zhong, Keith E. Latham — 2012-05-14

Objective: To examine whether spontaneous oocyte activation is determined by genetic differences and interacts with culture environment.Design: Experimental study.Setting: Temple University School of Medicine.Animal(s): C57BL/6, DBA/2, C3H/HeJ, and A/J strains, along with reciprocal F1 hybrid female mice (5–6 weeks).Intervention(s): Immature oocytes from different mouse strains collected and cultured in different maturation conditions, including different serum, serum replacement, bovine serum albumin (BSA), and follicle-stimulation hormone (FSH).Main Outcome Measure(s): The emission of first polar body, pronucleus formation, meiotic arrest, spontaneous activation, and expression of maturation regulators.Result(s): Oocytes from C57BL/6 mice display a high rate of delayed first meiotic division and spontaneous activation after the first meiotic division with in vitro maturation (IVM), and the second meiotic division with in vivo maturation (VVM) after superovulation. Spontaneous activation with IVM is sensitive to culture environment. Oocytes that spontaneously activated during the first meiotic division with IVM have unusual replicated sister chromatid pairs with slight connections at centromeres at first mitosis, whereas oocytes that activated in vivo display haploidization from the second meiotic division. Spontaneous activation is also seen in F1 hybrid oocytes, indicating a dominant trait from C57BL/6. Delayed meiosis was associated with reduced cyclin B and securin expression.Conclusion(s): Both mouse strain and culture environment have a statistically significant effect on the incidence of meiotic defects and spontaneous activation. Reduced expression of meiotic regulators may underlie this effect.

Studying nonobstructive azoospermia in cystinosis: histologic examination of testes and epididymis and sperm analysis in a Ctns−/− mouse model - Corrected Proof

2012-05-11

Objective: To study the pathogenesis of male infertility in cystinosis due to nonobstructive azoospermia, using a Ctns−/− mouse model.Design: Observational case-control study.Setting: Academic research laboratory.Animal(s): Male C57BL/6 Ctns−/− mice were compared with C57BL/6 wild-type (wt) mice.Intervention(s): None.Main Outcome Measure(s): Fertility was studied using litter size (n = 3 vs. n = 2). After animals were sacrificed, testes, epididymis, and vas deferens were removed for testicular cystine measurements (n = 5 vs. n = 6), histologic studies (n = 3 vs. n = 3), and sperm analysis (n = 3 vs. n = 3).Result(s): Mean testicular cystine content was significantly higher in Ctns−/− mice compared with wt mice (26.6 ± 1.22 vs. 0.1 ± 0.01 nmol cystine/mg protein). Testes of Ctns−/− mice had lower weight compared with wt mice (0.096 ± 0.009 g vs. 0.112 ± 0.004 g), but mice fertility was similar (litter size 6.6 ± 1.4 vs. 6.3 ± 2.6 pups). Neither histologic nor sperm abnormalities were found.Conclusion(s): The Ctns−/− mouse model generated on C57BL/6 background is not suitable for clarifying the pathogenesis of male infertility in cystinosis. The etiology of nonobstructive azoospermia in these patients remains unclear.

Human granulosa luteal cell oxidative phosphorylation function is not affected by age or ovarian response - Corrected Proof

2012-05-11

Objective: To safely prepare a functional autologous mitochondrial concentrate (MC) from follicular fluid (FF) cells, and to determine the effect of age and ovarian response on the oxidative phosphorylation (OXPHOS).Design: The nontoxicity of the MC was confirmed in human and mouse oocytes. The OXPHOS function was assessed by measuring the activity of succinate dehydrogenase (SDH) and cytochrome c oxidase (COX), and adenosine triphosphate (ATP) production in comparison with citrate synthase. The integrity of the mitochondrial DNA (mtDNA) was demonstrated by polymerase chain reaction (PCR).Setting: Tertiary hospital.Patient(s): A total of 40 patients undergoing IVF of heterogeneous ages and ovarian response.Animal(s): Superovulated 8- to 12-week-old female B6C3F1 mice.Intervention(s): None.Main Outcome Measure(s): A system for the preparation of functional nontoxic MC was established. The correlation between the mitochondrial mass and function to age and ovarian response was calculated. The integrity of mtDNA was demonstrated.Result(s): After injection into mouse oocytes, the MC did not interfere with parthenogenetic development. The MC OXPHOS function was intact. Total activity of SDH and COX was in correlation with the retrieved oocytes number, and in reverse correlation with age. However, after correction to the mitochondrial mass, COX and SDH activities were constant, unaffected by age or ovarian response. The mtDNA was intact in all samples, regardless of age and ovarian response.Conclusion(s): The function of the respiratory chain in mitochondria of FF cells is constant, unaffected by age or ovarian response.

Analysis of follicular fluid and serum markers of oxidative stress in women with infertility related to endometriosis - Corrected Proof

2012-05-11

Objective: To study the levels of four markers of oxidative stress in follicular fluid (FF) and plasma of patients with infertility related to endometriosis and controls.Design: Experimental study.Setting: University-affiliated hospital and infertility center.Patient(s): Ninety-one infertile women were included in the study (23 infertile women with endometriosis and 68 controls including infertile women due to tubal factor, male factor, or healthy egg donors).Intervention(s): Blood was obtained at the time of egg retrieval, and FF from the mature follicles of each ovary was centrifuged and frozen until analysis.Main Outcome Measure(s): Vitamin C and E, malondialdehyde, and superoxide dismutase concentrations in plasma and follicular fluid.Result(s): Women with endometriosis showed a lower vitamin C concentration in FF (12.7 ± 5.9 vs. 9.7 ± 6.9 μg/mL) and lower superoxide dismutase concentration in plasma (0.9 ± 1.4 vs. 0.5 ± 0.7 U/mL) compared with controls. Vitamin E plasma levels were significantly higher in women with endometriosis (8.1 ± 3.8 vs. 5.2 ± 3.2 μg/mL). A nonsignificant trend toward a lower plasma concentration of malondialdehyde was found in women with endometriosis.Conclusion(s): These findings suggest a lower antioxidant capacity in infertile women with endometriosis. Although a certain level of reactive oxygen species is required under physiological conditions, an altered balance between pro-oxidant and antioxidant activities may have an impact on folliculogenesis and adequate embryo development.

Effect of letrozole on estradiol production and P450 aromatase messenger RNA expression of cultured luteinized granulosa cells from women with and without endometriosis - Corrected Proof

Xiang Lu, Yu Wu, Xiao-Hong Gao, Yong-Wei Wang, Li Wang, Xiao-Xi Sun — 2012-05-11

Objective: To compare the estradiol production and P450 aromatase messenger RNA (mRNA) expression of cultured luteinized granulosa cells and the effect of letrozole on these parameters between women with and without endometriosis.Design: In vitro assays.Setting: Reproductive medical center.Patient(s): Patients undergoing in vitro fertilization (IVF): 23 patients with endometriosis and 19 controls without endometriosis.Intervention(s): Luteinized granulosa cells examined for estradiol levels and P450 aromatase mRNA expression in conditioned media at different letrozole concentrations of 0.0, 0.1, 1.0, and 10.0 μmol/L.Main Outcome Measure(s): Estradiol levels of the conditioned medium measured with an enzyme-linked immunosorbent assay (ELISA) kit and P450 aromatase mRNA expression determined by quantitative reverse transcription-polymerase chain reaction (QT RT-PCR).Result(s): The estradiol concentration of the conditioned media and P450 aromatase mRNA expression in the endometriosis group were statistically significantly lower than that of the control group, irrespective of letrozole concentrations. A statistically significant reduction of these two parameters was observed in both endometriosis and control groups at letrozole concentrations of 1 μmol/L and 10 μmol/L, but there were no statistically significant differences between letrozole concentrations of 1 μmol/L and 10 μmol/L. The number of high-quality embryos in the endometriosis group was statistically significantly lower than that of the control group.Conclusion(s): Lower estradiol production and P450 aromatase mRNA expression of cultured granulosa cells were found in women with endometriosis. Letrozole in the conditioned media further reduced these parameters.

Infertility, Diagnosis, Management, and IVF - Corrected Proof

John E. Buster — 2012-05-11

The title of this text is hardly imaginative. It does, however, describe exactly the content of this new text edited by Anil K. Dubey, Ph.D., a traditionally trained and scholarly basic scientist who is now IVF laboratory director at the George Washington University Reproductive Medicine Center. The text is a comprehensive assembly of contributions from 50 highly accomplished clinicians, basic scientists, administrators, and ethicists who have crafted their respective areas of expertise into 8 sections and 25 chapters. The work has a substantial international flavor, as its contributors comprise a diverse representation of European, Middle Eastern, Indian, and North American contributors.

Does ovulation induction with follicle-stimulating hormone still have a future in polycystic ovary syndrome? - Corrected Proof

Robert John Norman — 2012-05-09

Polycystic ovary syndrome (PCOS) is a common condition, regardless of the range of definitions available . Lack of regular ovulation is a cardinal component of the majority of presentations, with associated subfertility being a common reason for medical consultation. Initiation and successful conclusion of a pregnancy for women with PCOS in a timely, cost-effective, and safe manner is a skill required of all fertility specialists.

Epigenetics and vitrification of embryos: one step forward and many raised equivocal questions - Corrected Proof

Zeev Blumenfeld — 2012-05-07

In this issue of Fertility and Sterility, Zhao et al. have found that H19 putative imprinted control region (ICR) level of methylation significantly increased in vitrified two-cell bovine embryos and the generated blastocysts. They have also observed that incubation of vitrified-thawed 2-cell embryos with the DNA methyltransferase inhibitor 5-aza-2′-deoxycytidine (5-aza-dC) significantly reduced H19 ICR methylation and increased H19 expression by reverse transcription–polymerase chain reaction in the blastocysts derived from the vitrified embryos compared with those in fresh embryos.

Peritoneal cytokines and adhesion formation in endometriosis: an inverse association with vascular endothelial growth factor concentration - Corrected Proof

2012-04-30

Objective: To evaluate inflammatory/angiogenic cytokines—interleukin-1β (IL-1β), IL-6, IL-8, IL-12, interferon-γ (IFN-γ), tumor necrosis factor (TNF), and vascular endothelial growth factor A (VEGF-A)—in the peritoneal fluid of patients with endometriosis in relation to the occurrence and severity of pelvic adhesions and in control women without pelvic pathology.Design: Case-control study.Setting: University research institution and hospital.Patient(s): Sixty-five women with laparoscopically and histopathologically confirmed endometriosis, including 40 women with pelvic adhesions, and 37 control women without pelvic pathology.Intervention(s): Peritoneal fluid aspirated during routine diagnostic laparoscopic examination.Main Outcome Measure(s): Cytokines evaluated in the peritoneal fluid via specific enzyme-linked immunosorbent assays.Result(s): Endometriosis and the revised American Fertility Society score of this disease were associated with statistically significantly increased levels of peritoneal IL-6 and IL-8 whereas the incidence and score of endometriosis-related pelvic adhesions were negatively associated with increased levels of VEGF-A. Notably, the concentration of VEGF-A predicted adhesion development and severity after adjustment for endometriosis severity. The adhesion score also correlated with increased levels of IL-6; however, after adjustment for endometriosis severity, the effect of this cytokine was no longer statistically significant.Conclusion(s): Increased levels of VEGF-A may be associated with a decreased rate of pelvic adhesion formation in the course of endometriosis.

Uterus transplantation: animal research and human possibilities - Corrected Proof

Mats Brännström, Cesar Diaz-Garcia, Ash Hanafy, Michael Olausson, Andreas Tzakis — 2012-04-30

Uterus transplantation research has been conducted toward its introduction in the human as a treatment of absolute uterine-factor infertility, which is considered to be the last frontier to conquer for infertility research. In this review we describe the patient populations that may benefit from uterus transplantation. The animal research on uterus transplantation conducted during the past two decades is summarized, and we describe our views regarding a future research-based human attempt.

Antimüllerian hormone levels are independently related to ovarian hyperandrogenism and polycystic ovaries - Corrected Proof

2012-04-27

Objective: To determine the relationship of antimüllerian hormone (AMH) levels to polycystic ovaries and ovarian androgenic function.Design: Prospective case-control study.Setting: General clinical research center.Participant(s): Eumenorrheic asymptomatic volunteers without (V-NO; n = 19; reference population) or with (V-PCO; n = 28) a polycystic ovary and hyperandrogenemic anovulatory subjects grouped according to ovarian function into typical PCOS (PCOS-T; n = 37) and atypical PCOS (PCOS-A; n = 18).Intervention(s): Pelvic ultrasonography, short dexamethasone androgen-suppression test (SDAST), and GnRH agonist (GnRHag) test.Main Outcome Measure(s): Baseline AMH levels were related to polycystic ovary status, testosterone response to SDAST, and 17-hydroxyprogesterone response to GnRHag test.Result(s): AMH levels correlated with SDAST and GnRHag test outcomes. AMH was elevated (>6.2 ng/mL) in 32% of V-PCO versus 5% V-NO. The 21% of V-PCO who met Rotterdam PCOS criteria all had functional ovarian hyperandrogenism, but AMH levels were similar to nonhyperandrogenic V-PCO. AMH >10.7 ng/mL discriminated V-PCO from PCOS with 96% specificity and 41% sensitivity for PCOS-T, and insignificantly for PCOS-A.Conclusion(s): AMH levels are independently related to ovarian androgenic function and polycystic ovaries. Very high AMH levels are specific but insensitive for PCOS. In the absence of hyperandrogenism, moderate AMH elevation in women with normal-variant polycystic ovaries seems to indicate an enlarged oocyte pool.

Erratum - Corrected Proof

2012-04-26

In the article, “Sperm DNA damage measured by the alkaline Comet assay as an independent predictor of male infertility and in vitro fertilization success” (Fertil Steril 2011:95;652–7), the data provided in Table 3 under the column “Male Infertility” were interim data using just 15 of the 20 donors rather than using the complete group of 20 donors and 75 infertile males. The corrected values including all 20 donors are as in the amended table below. The authors regret the error.

Screening for partial AZFa microdeletions in the Y chromosome of infertile men: is it of clinical relevance? - Corrected Proof

2012-04-26

Objective: To evaluate the frequency of complete and partial AZFa Y-chromosome microdeletions among infertile Israeli men. To review the published frequencies and histologic findings of AZFa deletions.Design: Retrospective study.Setting: Academic medical center.Patient(s): A total of 1,260 infertile Israeli men. Literature review (2000–2010) of reports on men with AZFa deletions and their testicular findings.Intervention(s): The DNA of 1,260 infertile men was evaluated for AZF microdeletions. The DNA of 657 of them with undetected microdeletions was analyzed for partial AZFa deletion in the USP9Y and DDX3Y genes using sequence-tagged sites beyond EAA/EMQN recommendations.Main Outcome Measure(s): The frequency of complete and partial AZFa microdeletions. Availability of sperm cells for intracytoplasmic sperm injection in men with complete/partial microdeletions.Result(s): Two men had complete AZFa deletion (a frequency of 0.28% among nonobstructive azoospermic men). None had partial AZFa deletions.Conclusion(s): The likelihood of finding sperm cells in men with complete AZFa deletions is negligible. Complete AZFa deletion is rare and usually associated with azoospermia and absence of sperm cells in testicular tissue. The low frequency of partial AZFa deletions and the inconsistent prospects for spermatogenesis reported in the literature question the need for routine assessment of microdeletions in genes, such as USP9Y or DDX3Y.

Position on donors and smallpox vaccine: a committee opinion - Corrected Proof

2012-04-26

Although there is presently no definitive evidence linking vaccinia virus transmission through reproductive cells, SART/ASRM accordingly recommends that ART practitioners consider deferring donors who have recently received smallpox vaccine or contracted symptomatic vaccinia virus infection through close contact with a vaccine recipient (until after the vaccine or infectious scab has spontaneously separated). Good donor practice further suggests that donors who are not in good health, including those with recent complications from smallpox vaccine, should be similarly deferred. (This document was reviewed by the ASRM Practice Committee in 2011).

Recommendations for development of an emergency plan for in vitro fertilization programs: a committee opinion - Corrected Proof

2012-04-26

All in vitro fertilization (IVF) programs and clinics should have a plan to protect fresh and cryopreserved human tissue (embryos, oocytes, sperm) and to provide for continuation of patient care in the event of an emergency or natural disaster. This document was reviewed and affirmed by the Practice Committee in 2011.Earn online CME credit related to this document at www.asrm.org/elearn

Fertility treatment when the prognosis is very poor or futile: a committee opinion - Corrected Proof

The Ethics Committee of the American Society for Reproductive Medicine — 2012-04-26

The Ethics Committee recommends development of evidence-based policies that are patient-centered for each in vitro fertilization (IVF) center. In most cases, the provision of futile therapies is not ethically justiﬁable. For those treatments with very poor success rates, clinicians must be vigilant in their presentation of risks, beneﬁts, and alternatives. This document was reviewed in January 2012. This version replaces the previous version of this document, published in 2009.Earn online CME credit related to this document at www.asrm.org/elearn

Adverse perinatal outcome and in vitro fertilization singleton pregnancies: what lies beneath? Further evidence to support an underlying role of the modifiable hormonal milieu in in vitro fertilization stimulation - Corrected Proof

Suleena Kansal Kalra — 2012-04-26

The findings reported by Imudia et al. contribute to the emerging body of literature identifying modifiable aspects of IVF treatment that are associated with risk in singleton births. The most novel aspect of this study is that it presents a “threshold” effect of supraphysiologic E2 levels and subsequent antenatal and perinatal adverse outcome. In this study, outcomes were assessed in a group of patients undergoing IVF treatment and exposed to an elevated peak E2 level at a single institution, defined as an E2 level exceeding the 90th percentile on the day of hCG administration (>3,950 pg/mL), and compared to a group of patients undergoing IVF treatment with levels of E2 below that cutoff. Specifically, the study reports that the odds of preeclampsia was fourfold greater and small for gestational age infants was ninefold greater in patients undergoing IVF treatment with an elevated peak E2 level compared to women with a lower peak E2 level. No difference in preterm delivery was found.

Erratum - Corrected Proof

2012-04-26

In the article, “Is there a place for corifollitropin alfa in IVF/ICSI cycles? A systematic review and meta-analysis,” (Fertil Steril 2012;97:876–85), regarding OHSS incidence per woman randomized in four papers reviewed, the number needed to harm (NNT) was reported as 1, with an absolute risk increase of 1% after corifollitropin alfa. The correct NNT is 100. The authors regret the error.

Morphometric-stereological and functional epididymal alterations and a decrease in fertility in rats treated with finasteride and after a 30-day post-treatment recovery period - Corrected Proof

2012-04-23

Objective: To evaluate morphometric-stereological changes in the epididymal caput, sperm quality, and fertility parameters in rats treated with finasteride and after a 30-day post-treatment recovery period.Design: Experimental study in a research laboratory.Setting: Reproductive biology research laboratory.Animal(s): Male and female Sprague Dawley rats.Intervention(s): Treatment with finasteride (5 mg/kg/day) for 56 days followed by 30 days without treatment.Main Outcome Measure(s): Serum hormone analyses, morphometric-stereological and ultrastructural evaluation of the epididymal caput, sperm transit time, natural mating, in utero insemination, sperm membrane integrity, and fertility parameters.Result(s): Serum dihydrotestosterone levels in the finasteride group decreased by ∼40% compared with that of control rats. Ultrastructural analysis revealed significant reductions in several morphometric-stereological parameters of the epididymal caput. All parameters recovered significantly in the post-treatment period. There was no alteration in daily sperm production in the finasteride group. However, significant reductions in sperm transit time, motility, sperm membrane integrity, and fertility parameters were observed in rats treated with finasteride.Conclusion(s): Treatment with finasteride caused morphometric-stereological and functional changes in the epididymis and in sperm function that led to a reduction in fertility parameters. A 30-day post-treatment recovery period was insufficient to restore normal sperm motility, sperm transit time, and some fertility parameters.

Effect of laparoscopic excision of endometriomas on ovarian reserve: serial changes in the serum antimüllerian hormone levels - Corrected Proof

2012-04-23

Objective: To investigate the effect of laparoscopic endometrioma stripping on serum antimüllerian hormone (AMH) and the correlation between the clinicopathologic factors.Design: Prospective study.Setting: University hospital.Patient(s): Sixty-five women with endometriomas.Intervention(s): All patients underwent laparoscopic cystectomy. Serum AMH, FSH, LH, E2, and antral follicle count (AFC) were measured preoperatively, at 6 weeks, and at 6 months postoperatively. Specimens were analyzed histopathologically.Main Outcome Measure(s): The primary end point was to assess the ovarian reserve damage based on alterations of AMH and the secondary end point was to detect the changes in FSH, LH, E2, and AFC.Result(s): Serum AMH decreased significantly at the sixth month (61%) postoperatively. The FSH level increased significantly at the sixth week, but returned to normal at the sixth month. The AFC increased significantly at the sixth week and at the sixth month. The AMH level decrease was more evident in patients with the cyst <5 cm (65.7% vs. 41.3%). The AMH decrease was more in bilateral compared with unilateral endometriomas (67% versus 57%, respectively). No correlation was detected between the histopathologic analyses and tAMH level. Initially the AMH level was the only independent factor affecting the AMH decrease (odds ratio, 3.68; 95% confidence interval 1.66–8.14).Conclusion(s): Laparoscopic cystectomy of ovarian endometriomas causes a significant and progressive decline in serum AMH levels.

Activated glucocorticoid and eicosanoid pathways in endometriosis - Corrected Proof

2012-04-23

Objective: To define altered gene expression networks in endometriosis.Design: Experiments using endometriotic tissues and primary cells.Setting: Division of Reproductive Biology Research, Northwestern University.Patient(s): Premenopausal women.Intervention(s): Matched samples of eutopic endometrium and ovarian endometriosis (n = 8 patients) were analyzed by microarray and verified in a separate set of tissues (n = 6 patients). Experiments to define signaling pathways were performed in primary endometriotic stromal cells (n = 12 patients).Main Outcomes Measure(s): Using a genome-wide in vivo approach, we identified 1,366 differentially expressed genes and a new gene network favoring increased glucocorticoid levels and action in endometriosis.Result(s): Transcript and protein levels of 11β-hydroxysteroid dehydrogenase (HSD11B1), which produces cortisol, the biologically active glucocorticoid, were strikingly higher, whereas messenger RNA (mRNA) levels of the cortisol-degrading HSD11B2 enzyme were significantly lower in endometriotic tissue. Glucocorticoid receptor mRNA and protein levels were significantly higher in endometriosis. The inflammatory cytokine tumor necrosis factor robustly induced mRNA and protein levels of HSD11B1 and glucocorticoid receptor but suppressed HSD11B2 mRNA in primary endometriotic stromal cells, suggesting that tumor necrosis factor stimulates cortisol production and action. We also uncovered a subset of genes critical for prostaglandin synthesis and degradation, which favor high eicosanoid levels and activity in endometriosis.Conclusion(s): The proinflammatory milieu of the endometriotic lesion stimulates cortisol synthesis and action in endometriotic lesions.

Chromosomal abnormalities in embryos from couples with a previous aneuploid miscarriage - Corrected Proof

2012-04-23

Objective: To compare the incidence of chromosomal abnormalities in preimplantation embryos from couples undergoing preimplantation genetic screening (PGS) after previous aneuploid miscarriage after either natural conception (NC) or assisted reproductive technology (ART) versus fertile couples who underwent PGS for sex-linked diseases as a control group.Design: Retrospective study.Setting: IVF clinic.Patient(s): Patients with previous aneuploid conception undergoing PGS.Intervention(s): Embryo biopsy, fluorescence in situ hybridization.Main Outcome Measure(s): Embryo aneuploidy rates and pregnancy and implantation rates in couples with a previous aneuploidy for autosomes or sex chromosomes.Result(s): The overall rates of chromosomal abnormalities in groups with previous autosomal aneuploidy were significantly higher compared with the control group (67.8% for those whose previous aneuploidy arose after NC and 65.8% for those previously arising after ART, vs. 34.0%). No significant differences were observed in those with previous sex chromosome abnormalities compared with control subjects. Within couples with previous aneuploidies after NC, no difference existed in the incidence of chromosomal abnormalities compared with the ART groups. Clinical outcomes were better (trend) in patients with previous autosomal aneuploidy after NC.Conclusion(s): In preimplantation embryos, the incidence of chromosomal abnormalities due to a previous aneuploid miscarriage after either NC or ART is significantly higher than in the control group. Furthermore, this incidence is higher when the previous aneuploidy was for autosomes; PGS is recommended in these couples.

The association between fertility clinic performance and cycle volume: implications for public reporting of provider performance data - Corrected Proof

Dan Gong, Emre Seli — 2012-04-23

Objective: To quantitatively determine the relationship between fertility clinic performance and cycle volume, and to assess the implications for public reporting of provider performance data.Design: Retrospective longitudinal analysis.Setting: Clinic.Patient(s): The study population included 307 U.S. assisted reproductive technology (ART) clinics that continuously reported performance data to the Centers for Disease Control from 2003 to 2008.Intervention(s): None.Main Outcome Measure(s): Regression coefficients between pregnancy rate per cycle, live birth rate per cycle, or live birth rate per transfer and number of ART cycles performed.Result(s): Overall, there was no association found between a clinic's most recently reported success rate and its cycle volume. This finding was consistent across three time periods studied. Moreover, stratification analyses of clinics with greater than ±5%, ±10%, and ±20% change in success rates also found no association between clinic performance and cycle volume.Conclusion(s): As proxied by cycle volume data, patients seeking ART treatment do not seem to be influenced by positive or negative changes in a clinic's performance despite the public availability of this data. These results suggest that current public quality reporting encompassing success rates alone will not change patient behavior and therefore is insufficient to place salient competitive pressure on health care providers. Further research is necessary to define provider performance comprehensively and to determine the metrics, if any, to which patients respond.

Role of routine monitoring of liver and renal function during treatment of ectopic pregnancies with single-dose methotrexate protocol - Corrected Proof

2012-04-23

Objective: To assess whether or not baseline serum transaminases and creatinine measurements, before administration of methotrexate, identified significant liver or kidney disease, which have the potential to alter the management plan for the treatment of ectopic pregnancies.Design: This is a retrospective study of patients treated for ectopic pregnancy.Setting: Women's emergency room and reproductive endocrinology office at a teaching hospital over a 3-year period.Patient(s): Women presenting for treatment of ectopic pregnancy.Intervention(s): None.Main Outcome Measure(s): Assessment of baseline serum transaminases and creatinine measurements before administration of methotrexate to identify significant liver or kidney disease.Result(s): A total of 383 patients were managed for ectopic pregnancy from January 2006 to December 2008. Of these, 320 patients received methotrexate as part of their treatment. No patient was denied treatment with methotrexate secondary to concerns regarding liver or renal function. No complication related to methotrexate administration was documented. A subgroup of 81 patients had pre- and postadministration labs, and no significant difference was noted upon comparing the values.Conclusion(s): Routine measurement of serum aspartate aminotransferase and creatinine levels may not be necessary before instituting a single-dose methotrexate treatment regimen for the management of ectopic pregnancy.

Running interference - Corrected Proof

Nanette Santoro — 2012-04-23

Carpenter et al. report on secondary data from the MsFlash randomized, controlled trial of escitalopram for hot flashes in menopausal women. The trial demonstrated a statistically significant reduction in hot flash frequency versus placebo at 8 weeks in the primary trial . Hot flash severity decreased in a similar direction and magnitude to frequency. The study demonstrated a marked placebo effect, with a decrease in hot flash frequency of 3 per day by 8 weeks in the women taking placebo. The treatment effect was slightly more, 4.6 hot flashes per day.